AllPrep DNA/RNA/Protein Mini Kit

For simultaneous purification of DNA, RNA, and protein from cells and tissues

Maximal recovery of each analyte from precious samples

Time savings through short, streamlined workflow

Comparable data as analytes come from same sample

No need for organic solvents such as phenol and acetone

The AllPrep DNA/RNA/Protein Mini Kit allows simultaneous purification of DNA, RNA, and protein from the same cell or tissue sample. The kit can be used in combination with Allprotect Tissue Reagent, which delivers immediate stabilization of DNA, RNA, and protein in tissue samples for long-term storage without freezing. Tissues can be efficiently disrupted using a TissueRuptor or TissueLyser system.

Rat tissues were stabilized in Allprotect Reagent, and DNA and RNA were purified using the AllPrep DNA/RNA/Protein Mini Kit. The CT values in real-time PCR and real-time RT-PCR analyses were similar to those achieved with tissues stabilized in liquid nitrogen.||Rat tissues were stabilized in Allprotect Reagent, and protein was purified using the AllPrep DNA/RNA/Protein Mini Kit. Duplicates were run on an SDS-PAGE gel, followed by [A] western blotting for ERK2 and [B] Coomassie staining.|

The AllPrep DNA/RNA/Protein Mini Kit standardizes sample preparation for systems biology. DNA, RNA, and protein are all prepared from the same source, eliminating the variation inherent in preparing these analytes from different samples.There is no need to split the sample into 3 prior to purification, allowing maximum recovery of DNA, RNA, and protein. In addition, no organic solvents such as phenol and acetone are required.

Procedure

Cells or tissues are first lysed and homogenized in Buffer RLT, which immediately inactivates DNases and RNases as well as proteases to ensure isolation of intact DNA, RNA, and proteins. The lysate is then passed through an AllPrep DNA spin column. This column, in combination with the high-salt buffer, allows selective and efficient binding of genomic DNA. The column is washed and pure, ready-to-use DNA is then eluted.

Ethanol is added to the flow-through from the AllPrep DNA spin column to provide appropriate binding conditions for RNA, and the sample is then applied to an RNeasy spin column, where total RNA binds to the membrane and contaminants are efficiently washed away. High-quality RNA is then eluted in RNase-free water.

Buffer APP, a novel aqueous protein precipitation solution, is added to the flow-through of the RNeasy spin column, and the precipitated proteins are pelleted by centrifugation. Intact total proteins are redissolved in an appropriate buffer and then ready to use in downstream applications. The kit includes Buffer ALO, which is compatible with SDS-PAGE, for dissolving the protein pellet. (See flowchart "AllPrep DNA/RNA/Protein procedure".)

Applications

The AllPrep DNA/RNA/Protein Mini Kit streamlines sample preparation by enabling simultaneous purification of genomic DNA, total RNA, and total protein from the same cell or tissue sample. Total protein purified using the kit is suitable for SDS-PAGE and western blotting. Genomic DNA and total RNA purified using the AllPrep DNA/RNA/Protein Mini Kit deliver optimal results in all downstream applications including: